Shin et al.

Bioresources and Bioprocessing (2025) 12:3 Bioresources and Bioprocessing

https://doi.org/10.1186/s40643-024-00836-7

REVIEW Open Access

Review on mushroom mycelium‑based

products and their production process:
from upstream to downstream
Hyun‑Jae Shin1* , Hyeon‑Su Ro2, Moriyuki Kawauchi3 and Yoichi Honda4*

Abstract
The global trend toward carbon neutrality and sustainability calls for collaborative efforts in both the basic
and applied research sectors to utilize mushroom mycelia as environmentally friendly and sustainable materi‑
als. Fungi, along with animals and plants, are one of the major eukaryotic life forms. They have long been utilized
in traditional biotechnology sectors, such as food fermentation, antibiotic production, and industrial enzyme
production. Some fungi have also been consumed as major food crops, such as the fruiting bodies of various
mushrooms. Recently, new trends have emerged, shifting from traditional applications towards the innovative
use of mushroom mycelium as eco-friendly bioresources. This approach has gained attention in the development
of alternative meats, mycofabrication of biocomposites, and production of mycelial leather and fabrics. These
applications aim to replace animal husbandry and recycle agricultural waste for use in construction and electrical
materials. This paper reviews current research trends on industrial applications of mushroom mycelia, covering
strain improvements and molecular breeding as well as mycelial products and the production processes. Key find‑
ings, practical considerations, and valorization are also discussed.
Keywords Mushroom, Mycelium, Biomaterials, Biocomposite, Mycofabrication, Upstream, Downstream, Molecular
breeding, Valorization, Bioproducts

*Correspondence:
Hyun‑Jae Shin
[email protected]
Yoichi Honda
[email protected]
Full list of author information is available at the end of the article

© The Author(s) 2025. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which
permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the
original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or
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licence, visit http://​creat​iveco​mmons.​org/​licen​ses/​by/4.​0/.
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 2 of 21

Graphical Abstract

Introduction resources, and pollutes air, soil, and water bodies dur-
Recently, worldwide awareness of animal welfare and ing production, transportation, and demolition. Eight
environmental protection has been increasing, and to ten percent of global carbon dioxide emissions are
interest in vegan alternatives and decreased plastic use released from manufacturing construction materials,
is rising. In particular, there is a movement to substitute and plastic bans have been registered in many countries
plant materials for leather and meat, mimicking animal (Rathore et al. 2021).
materials (Rollin 2006). Animal husbandry is respon- Fungi belong to a major group of eukaryotic organ-
sible for 15% of total greenhouse gas emissions devas- isms. They have a long evolutionary history since their
tating land and water environments. The generation of emergence about 1.2–1.5 billion years ago (Wang et al.
odors and wastewater during the manufacturing pro- 1999). Among various fungal groups, saprotrophic
cess of meats and leathers is one of the leading causes fungi belonging to the Agaricomycetes class are attrib-
of environmental pollution (Covington and Wise 2020). uted to the decline of terrestrial coal deposition at the
Accordingly, research on alternative materials and eco- early Permian period (300 million years ago) (East-
friendly processes using renewable biomass is actively wood 2014; Hawksworth 2012; Floudas et al. 2012),
being conducted to reduce environmental pollution although tectonic activities and climate conditions
and pursue a healthy lifestyle (Lazar et al. 2023). Most may also contribute to the carbon burial (Nelson et al.
industrially fabricated materials, such as construction 2016). Among these, 155,603 species have been docu-
and packaging materials nowadays, are nonrecyclable mented, which represents less than 10% of the esti-
and environmentally unfriendly. Utilizing these con- mated 2.5 million fungal species (Niskanen et al. 2023).
ventional materials consumes energy, limits natural Many fungal members have been used in the traditional
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 3 of 21

biotechnology sectors, such as food fermentation, electrical, and industrial materials (Ferrand 2024; Ala-
antibiotics, and industrial enzyme production. Some neme et al. 2023; Danninger et al. 2022). Although many
saprotrophic fungi are cultivated as major food crops, review articles and research papers have been published
such as fruiting bodies of various mushrooms. ’Mush- on mycelium-based materials, there has been no review
room’ refers to fungal species that can produce large- of the vertical approach from mushroom strains and
sized fruiting bodies that are observable to the naked mutation methods as biological resources to the myce-
eye, including the mycelial network and fruiting body lium production process. Therefore, this review discusses
itself. Among mushroom-forming fungi, wood-decay- comprehensive studies from upstream (strain selection
ing fungi have long been a subject of fascination for and breeding) to downstream (product development and
researchers, with their potential applications ranging process design) processing. Readers of this paper will
from culinary delights to remediation of environmen- have a more in-depth understanding of recent research
tal pollutants. Hereafter, vegetative mycelia and fruit- on mushroom mycelium research and its limitations.
ing bodies of wood-decaying Agaricomycete fungi are
the focus of interest for environmentally friendly and Fungi and mushrooms as bioresources
sustainable materials for the future. In recent years, a Wood‑decay fungi
dramatic increase in global mushroom fruiting body Fungi, with their ancestral roots in aquatic habitats fea-
production has been driven partly by the growing turing flagellated spores, exhibit a closer evolution-
awareness of their nutritional and therapeutic benefits ary affinity to animals than plants. (James et al. 2006).
(Bhagarathi et al. 2023). More recently, new trends have Nonetheless, most of them have diversified by interact-
been shifting away from traditional biotechnology and ing with plants throughout evolution, colonizing terres-
applying fungal mycelia as eco-friendly resources in trial environments. They coexist with plants, decompose
the formulation of alternative meats, fabrication of bio- dead plant matter, or live parasitically on plants. There
composites, and production of mycelial leather, aiming are symbiotic mushrooms like mycorrhiza that cannot
to replace animal husbandry and recycle agricultural degrade wood. Only certain Agaricomycetes are able to
wastes (Madusanka et al. 2024; Aiduang et al. 2024; degrade wood efficiently, called wood-decaying fungi.
Yang et al. 2021; Antinori et al. 2020; Kim et al. 2017; Based on their decomposition abilities, they are roughly
Holt et al. 2012). classified as white-rot fungi and brown-rot fungi.
Mycelium is a fast-growing vegetative section of a Brown rot fungi can decompose cellulose but cannot
fungus that is commonly found in biological and agri- fully break down lignin. They produce hydroxyl radicals
cultural wastes. It is a harmless and sustainable material generated from the redox reaction between ­H2O2 and
that bonds with the media to which it is attached. The ­Fe3+ ions through the Fenton reaction, oxidizing the crys-
Material Innovation Initiative, a non-profit think tank, talline cellulose and lignin in wood and creating cracks in
has classified non-animal-based materials, excluding ani- the wood structure (Arantes and Goodell 2014). As the
mal leather, fur, wool, down, and silk, as next-generation hyphae grow through these cracks, they secrete hemicel-
materials (materialinnovation.org 2022). To be classi- lulase and endoglucanase enzymes to further decompose
fied as a next-generation material, an item must have a wood components such as hemicellulose and cellulose
low negative environmental impact throughout its life (Arantes and Goodell 2014; Krah et al. 2018). Conse-
cycle, including extraction, production, and consump- quently, wood degradation by these fungi leaves a brown-
tion. Examples of next-generation materials include ish cubical fragmentation pattern on the wood. Fungal
plant materials, fungal mycelia, cell cultures, microbes, species such as Coniophora puteana, Fomitopsis pini-
and recycled materials. Based on the above definition, cola, Gloeophyllum trabeum, Postia placenta, Serpula
mushroom (mycelium) derived novel materials are next lacrymans, and Wolfiporia cocos, are some of the known
generation eco-friendly materials. Mycelium-based vegan brown rots (Floudas et al. 2012).
leather and packaging are created by growing mycelia White-rot fungi are saprotrophs capable of completely
into customized growth chambers and molds, so-called decomposing lignin through the activity of oxidative
mycofabrication (Raman et al. 2022). The field of myce- enzymes, such as laccase and lignin-modifying peroxi-
lium-based leather and packaging has been expanding dases (PODs), leaving behind white, cellulosic, and hemi-
(Ariyani et al. 2024; Kniep et al. 2024; Madusanka et al. cellulosic polymers in decaying wood (Cui et al. 2021).
2024; Amobonye et al. 2023; Raman et al. 2022). Patents These polymers are further decomposed by cellulase and
in this area including various manufacturing methods are hemicellulase. White-rot fungi are the primary degrad-
also abundant and publicly available (Elsacker et al. 2023). ers in forests, and many species among them, such as
Beyond vegan leather/textile and packaging, mushroom Agaricus bisporus, Lentinula edodes, Pleurotus ostrea-
mycelium-based materials expand into construction, tus, Pl. eryngii, and Flammulina velutipes, are consumed
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 4 of 21

Fig. 1 Fungal cell wall structure and its morphology. Cell wall is mainly composed of β-glucan, chitin, and proteins. The model is based on recent
cell wall analysis data from Sc. commune

as edible mushroom. White-rot fungi generally possess three-dimensional mycelial network. The three-dimen-
multiple copies of genes for laccase, PODs, cellulase com- sional structure of mycelium is a complex and intercon-
plex, and glucoside hydrolases (GHs) through paralogous nected network of fungal filaments (Islam et al. 2017).
gene expansion (Floudas et al. 2012). The following spe- Mycelial strength is mainly attributed to cell wall poly-
cies are some of the known white-rot fungi: Auricularia saccharides (Gow et al. 2017). The fungal cell walls in
delicata, Fomitiporia mediterranea, Ganoderma luci- higher fungi (fungi belonging to Ascomycota and Basidi-
dum, Heterobasidion annosum, Perenniporia fraxinea, omycota) consist of 10–20% chitin, 50–60% glucans, and
Pl. ostreatus, Punctularia strigosozonata, Schizophyllum 20–30% glycoproteins (Feofilova 2010; Bowman and Free
commune, Stereum hirsutum, and Trametes versicolor. 2006).
Polyporales are saprotrophic or pathogenic mushroom The cell wall components of mushrooms play a crucial
species that produce shelf- or bracket-shaped fruiting role in determining their material properties and electri-
bodies around tree trunks or branches. These fungi cre- cal characteristics. Comparing these aspects can provide
ate strong mycelia as they attach to wood and produce insights into the significance of genetic modifications
sturdy, woody fruiting bodies. Bae et al. (2021) observed affecting mushroom cell walls (Fig. 1).
mycelial growth by cultivating 64 strains of Polyporales Chitin A long-chain polymer of N-acetylglucosamine
fungi, including Tr. versicolor, Pycnoporus coccineus, Ga. that provides mechanical strength and rigidity to the cell
lucidum, and Pe. fraxinea, on sawdust and liquid media. wall (Ehren et al. 2020).
Their findings revealed significant variation in growth β-glucans Polysaccharides that contribute to the overall
rates at the strain level, with Pe. fraxinea exhibiting the structure and flexibility of the cell wall (Ehren et al. 2020).
fastest growth. Scanning electron microscope (SEM) Proteins Present in the outer layer of the cell wall, pro-
analysis of the mycelial mat suggested that even among teins like mannoproteins and hydrophobins contribute
Polyporales strains, there were notable differences in to cell wall integrity and hydrophobicity (Haneef et al.
both mycelial density and hyphal diameter, which may 2017).
correlate with the strength of mycelial mat, demonstrat- The cell wall structure of the Agaricomycete Sc. com-
ing the necessity for further exploration and isolation of mune has previously been analyzed (Ehren et al., 2020,
diverse strains. Safeer et al. 2023). Agaricomycete cell walls are assumed
to consist of rigid chitin-glucan complexes in the inner
Fungal cell wall cell wall and a more flexible network of β-glucans, extra-
Fungal mycelia is made of rigid tubes or cylinders of cellular polymeric matrix (EPS) and other proteins in the
connected cells that grow on the surface of and inside outer cell wall (Fig. 1) (De Beeck et al. 2021, Ehren et al.
solid substrates and produce aerial hyphae, forming a 2020, Safeer et al. 2023). The inner cell wall is made of
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 5 of 21

different forms of chitin from single stranded to highly These terms, in turn, relate to the functions and struc-
branched, as well as β-(1,3), β-(1,3)-(1,6)-glucan, and tural characteristics of three main types of hyphae: gen-
α-(1,3)-glucan (Ehren et al. 2020). erative, binding, and skeletal hyphae (Fig. 2). Monomitic
Chitin is a homopolymer of N-acetylglucosamine hyphae consist of exclusively generative hyphae, and are
linked through β-1,4 glycosidic bonds catalyzed by mem- often saprotrophic, growing in softer environments like
brane-bound chitin synthases (CHSs). Fungi have mul- soil or decaying organic matter (e.g., Ag. bisporus). Dim-
tiple copies of CHS genes classified into seven classes of itic hyphae are made of either generative and skeletal, or
CHS, belonging to two structurally distinct families (Ron- generative and binding hyphae (e.g., Grifola frondosa),
cero 2002; Lenardon et al. 2010). The number of CHS whereas trimitic hyphae consist of all three types: gen-
genes generally correlates with the cell wall chitin con- erative, skeletal, and binding hyphae (e.g., Tr. versicolor).
tent (Latgé and Calderone 2006). Disruption of CHSs in Dimitic and trimitic fungi are more specialized for ligno-
Aspergillus fumigatus causes increased sensitivity to cell cellulosic substrates like wood and often exhibit higher
wall perturbing agents (Muszkieta et al. 2014). Deletion resistance to environmental stress, making them eco-
of CSMA or CSMB, belonging to Family II CHS, shortens logically important in forest ecosystems and valuable
chitin fibrils resulting in swollen conidia and reduced vir- for applications such as biodegradation and bioremedia-
ulence (Muszkieta et al. 2014). In pathogenic yeast Can- tion. Nowadays, many mycelium-oriented companies use
dida albicans, the chs8 mutant showed a defect in the Ganoderma species for their products because of their
synthesis of long-chitin microfibrils, which were found at trimitic hyphae structure (Porter and Naleway 2022).
the bud scar and septa, whereas the chs3 mutant exhib-
ited a defect in the accumulation of short-chitin rodlets Mushroom strain improvement
as a cell wall component (Lenardon et al. 2007; Gow and Purpose of strain improvements
Lenardon 2023). In the agaricomycete Pl. ostreatus dis- Various fungal species including Polyporales, Fomes
ruption of basidiomycete-specific chitin synthases chsb2, fomentarius, Pl. eryngii, Tr. versicolor, and Agaricus have
chsb3 and chsb4 causes increased sensitivity to cell wall been explored for their potential in creating sustainable
perturbing agents, thinner cell walls and decreased aerial leather alternatives (Kniep et al. 2024). To improve the
hyphae production (Schiphof et al. 2024). fundamental properties of mycelium-based materials,
The glucan components are polymers of glucose units mycelium composites and/or mycelial mats with novel
linked through β-1,3, β-1,6, α-1,3, and α-1,4 glycosidic functions and characteristics can be developed by molec-
bonds, with β-1,3-glucan being the main cell wall con- ular breeding of fungi. With a shorter life cycle, mush-
stituent (Beauvais et al. 2001; Papaspyridi et al. 2018; Liu room-forming fungi have great potential to be developed
et al. 2022a, b; Bowman & Free 2006). The synthesis of as a new cultivar with a wider variety of properties com-
β-1,3-glucan is catalyzed by the membrane-integrated pared to other material crops and farm animals. Many
glucan synthase, Fks (Hu et al. 2023). Agaricomycete technologies such as chemical, physical, and molecular
have fewer diverse fks genes than chs genes as they have biological protocols have been introduced and success-
2–4 fks and 8–9 chs copies (Nakazawa et al. 2024; Grigo- fully developed in some mushroom-forming fungi. This
riev et al. 2014). Cell wall proteins are linked to chitin or facilitates faster and easier molecular breeding to develop
glucans through charged amino acids (Safeer et al. 2023). desired properties for future requirements such as food,
EPS is a loosely packed matrix of mucilage that covers the medicine and sustainable biomaterials (Salazar-Cerezo
cell wall. It comprises polymeric materials like polysac- et al. 2023; Dong et al. 2022; Lee et al. 2011; Sathesh-
charides and glycoproteins (De Beeck et al. 2021; Gow Prabu and Lee 2011). Moreover, their relatively small
et al. 2017). genome size makes it easier to check the whole genome
sequence of isolated strains, which may ensure foreign-
Hyphae type DNA-free edited cultivars are not classified as genetically
As the mycelium is made up of hyphae, mycelium physi- modified organisms (GMOs) (Nakazawa et al. 2024). In
ochemical properties of the mycelium depend on those the following sections, these breeding strategies will be
of the hyphae. Therefore, identifying the shape and char- introduced and discussed in more detail.
acteristics of the mycelium is one of the most important Generally, chemical and physical treatments were used
factors in the selection of mushrooms as a biological to introduce random mutations and isolate a mutant
resource (Alaneme et al. 2023; Nadeem and Pirzada 2018; strain with a suitable phenotype for breeding. In genetic
Montes et al. 2003). The terms monomitic, dimitic, and breeding, a mating cross of two different strains is used
trimitic are used in mycology to classify fungi based on to select favored progenies with desired phenotypes. In
the types of hyphae present in their fruiting bodies and contrast, molecular breeding methods directly intro-
mycelia (Jones et al. 2020; Porter and Naleway 2022). duce a mutation in a targeted gene, or heterogeneous
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 6 of 21

Generative hyphae Binding hyphae Skeletal hyphae

- Branched
- Branched - Unbranched
- Thin-walled
- Thick-walled
- Septate - Thick-walled
- Interwoven the other
- Clamp connection
hyphae

<Monomitic> <Dimitic> <Trimitic>

Fig. 2 Classification of hyphae type and its mushroom example

recombinant gene, in the cell, using genetic transforma- method is particularly useful for generating resources
tion and/or genome editing. to advance genetic research and deepen our under-
standing of the underlying mechanisms behind muta-
Chemical treatments tion-induced traits.
The use of chemical reagents to mutate mushroom In the case of Ganoderma lingzhi, chemical muta-
strains is an established technique in mycelium gens like lithium chloride have been used to enhance
research, aimed at improving mushroom breeding the production of bioactive compounds such as poly-
and production characteristics. Chemical mutagenesis saccharides and triterpenoids. This approach resulted
involves the use of chemical agents to induce mutations in significant increases in yield compared to the origi-
in the genetic material of organisms, leading to the nal strains, demonstrating the potential of chemical
development of new strains with potentially desirable mutagenesis in improving the commercial viability
traits. Chemical mutagens such as methanesulfonate of mushroom strains (Ma et al. 2018). After inducing
methylester, an alkylating agent, have been used to mutations, careful screening and selection of the
treat basidiospores of mushrooms like Hypsizygus mar- desired mutant strains is crucial. This process often
moreus. This treatment can yield mutant monokaryotic involves assessing morphological changes and growth
mycelia, which are then selected and mated to produce characteristics to identify strains with beneficial traits.
dikaryons with improved characteristics. For instance, One of the advantages of using chemical mutagenesis
some mutant strains have shown increased fruiting is that the resulting strains are typically classified as
body production, while others exhibited unique mor- non-transgenic. This classification can be beneficial in
phological features such as flattened stipes and pilei markets where non-genetically modified organisms are
(Lee et al. 2011). Chemical mutagenesis can lead to the preferred. Overall, chemical mutagenesis is a power-
development of strains that exceed parental cultivation ful tool in mushroom strain development, offering the
characteristics, making them valuable for both com- potential to enhance yield, improve growth character-
mercial cultivation and molecular genetic studies. This istics, and increase the production of valuable bioactive
compounds.
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 7 of 21

Ultraviolet (UV) and gamma (γ) ray irradiation that it does not require a protocol for efficient protoplast
Ultraviolet (UV) irradiation is widely used to induce production (de Groot et al. 1998; Kim et al. 2015). How-
DNA mutation in mushrooms due to its effectiveness ever, the ATMT method does not yield large numbers of
in generating genetic diversity and enhancing desirable transformants in a single experiment, and the introduced
traits (Sathesh-Prabu and Lee 2011). UV light causes DNA almost always integrates on the chromosome, clas-
DNA damage, leading to mutations which can result sifying resulting strains as GMOs. Furthermore, the
in new phenotypes (Dong et al 2022). This method has ATMT method cannot directly introduce proteins into
been successfully applied to various mushroom species to the cell.
improve traits such as yield, tolerance to environmental Gene targeting technology can be used to disrupt or
stress, and biosynthetic capabilities (Sun et al. 2022a, b; introduce a desired change in a gene of interest. When
Harfi et al. 2021; Wang et al. 2014). UV irradiation has a gene is disrupted in this manner it is referred to as a
not yet been applied directly to mycelia for the purpose gene knock-out However, developing gene-targeting
of developing mycelium-based materials. The effect of technology is challenging because the homologous
UV treatment on multinucleate hyphae remains an area recombination system is active only during meiotic divi-
worthy of further study. In particular, gamma irradiation sion just before basidiospore formation and insufficiently
has been suggested as a useful alternative to overcome active in the vegetative mycelia or during isolation of
the limitations of hybridization and the environmental transformants from protoplasts. Non-homologous end
and food safety concerns of genetically modified crops joining (NHEJ) is the other repair system, predominant
(Riviello-Flores et al. 2022). It can mutate fungal growth through most of the fungal life cycle. In a limited number
stages and mycelium properties to select targeted or use- of model species such as Coprinopsis cinerea (Nakazawa
ful variants, and is rapid and stable compared to other et al. 2011), Sc. commune (Jan Vonk et al. 2019) and Pl.
breeding methods (Sathesh-Prabu and Lee 2011). By irra- ostreatus (Salame et al. 2012), specific genes required for
diating monokaryon spores with gamma rays at appro- NHEJ system were disrupted to develop a strain with an
priate doses ranging from 50 to 200 Gy, researchers have efficient homologous recombination system.
successfully induced mutations that result in diverse Conventional genetic transformation methods typi-
dikaryon mycelia (Kim and Yu 2014). Additionally, the cally involve the presence of foreign DNA sequences,
analysis of genetic variation in different mushroom due to the use of a genetic marker gene for the screen-
strains, such as Hy. marmoreus and Lyophyllum species, ing of transformants. In most cases, genetic markers con-
using techniques like random amplification of polymor- tain recombinant DNA composed of multiple origins of
phic DNA (RAPD) has demonstrated the effectiveness organisms (Matsunaga et al. 2017). Limited numbers of
of gamma-ray irradiation in developing new mushroom genetic markers are known as a ‘self-cloning marker’, such
strains with enhanced traits (Kim and Yu 2014). These as pyrG (Nakazawa et al. 2016) or CbxR markers (Honda
findings highlight gamma irradiation as a crucial strategy et al. 2000). It must be considered which kind of mark-
for mushroom mutation, enabling the breeding of novel ers or what kind of purposes the conventional transfor-
mushroom varieties with improved characteristics. Some mation must be considered when developing new strains
mutant strains, including Le. edodes, Ga. lucidium, and aimed at future development of mycelial composites or
Sc. commune showed good hyphae growth rate and den- food productions.
sity on saw-dust media (Kim et al. 2020). In these experi-
ments, a number of variables need to be determined, Genome editing
including the effect of gamma-irradiation on genes and Genome editing is a promising technology for molecu-
the propensity of the mutant strain to revert to the origi- lar breeding in many organisms including mushroom-
nal parent strain. forming fungi. The most popular and efficient genome
editing technique, CRISPR/Cas9 (Jinek et al. 2012; Song
Genetic transformation et al. 2019) was successfully introduced into various
A conventional genetic transformation system is a core mushroom-forming fungi including Cop. cinerea (Sug-
technology for introducing recombinant DNA or pro- ano et al. 2017), Sc. commune (Jan Vonk et al. 2019), Pl.
teins into living cells and establishing new strains with ostreatus (Boontawon et al. 2021b; Xu et al. 2022; Yama-
desired phenotypes based on the introduced genetic fac- saki et al. 2022; Koshi et al. 2022), Le. edodes (Moon
tors. Two approaches have been used majorly to intro- et al. 2021; Kamiya et al. 2023), Ga. lucidum (Qin et al.
duce recombinant DNA into the fungal cell: one depends 2017; Liu et al. 2020), Ag. bisporus (Choi et al. 2023), Pl.
on protoplast, and the other on a special kind of bacte- eryngii (Wang et al. 2021), Fl. filiformis (Liu et al. 2022a,
rium, Agrobacterium tumefaciens. Ag. tumefaciens-medi- b), and Gelatoporia (formally, Ceriporiopsis) subver-
ated transformation (ATMT) method has an advantage in mispora (Nakazawa et al. 2022). CRISPR/Cas9 requires
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 8 of 21

two essential components: a Cas9 protein that intro- surface, guanine nucleotide exchange factors (GEFs), Rho
duces DNA double-strand break (DSB) and a guide RNA GTPases, protein kinase C (PKC) and a mitogen-acti-
(gRNA) that recruits Cas9/gRNA complex to the target vated protein kinase (MPK) module (Dichtl et al. 2016).
site on the chromosome through base pair formation In basidiomycetes, the CWI signaling pathway in
with its corresponding single stranded DNA. Gener- Cryptococcus neoformans has been well studied. In Cr.
ally, the introduced DSB is repaired mainly by the NHEJ neoformans, the major components of the CWI signal-
repair system in the cell. However, small insertions or ing pathway were conserved, with the exception of the
deletions of base pairs may happen at a certain frequency membrane-anchored sensor proteins (Dichtl et al. 2016).
during the repair process. In a case where no mutation is In the yeast-like basidiomycetes Ustilago maydis and Cr.
introduced, the target site would be attacked repeatedly neoformans, Rho1 is essential for growth (Dichtl et al.
by the Cas9/gRNA complex until a mutation occurs. In 2016). In U. maydis, the necessity of Rho1 for cell divi-
this way, when using a gRNA sequence specific to the tar- sion and chitin deposition was demonstrated using a
get site, CRISPR/Cas9 can be used to disrupt a gene of strain where Rho1 driven by the crg1 promoter (Pham
interest (Boontawon et al. 2021a; Nakazawa et al. 2023). et al. 2009). However, in Ga. lucidum, downregulation
With the co-transformation donor DNA consisting of of slt2/mpk1 by RNAi resulted in a moderately strong
DNA fragments flanked by homologous arm sequences growth defect on both agar and substrate medium
(HR arms) to a target gene, CRISPR/Cas9 can efficiently (Zhang et al. 2017). In this strain, both the amount of
catalyze the exchange of DNA sequences between the glucan and chitin in the cell wall and the expression of
chromosome and donor DNA through homology-direct the corresponding biosynthetic genes were reduced.
repair (HDR) at the target site (Jan Vonk et al. 2019). The Thus, in basidiomycetes, MPK1 has a strong effect on
efficiency of gene editing with CRISPR/Cas9 combined growth while allowing modification of cell wall compo-
with HR arms is generally greater than with CRISPR/ sition, suggesting that downstream transcription factors
Cas9 alone; however, the effectiveness can vary depend- are promising breeding targets. Indeed, in Ga. lucidum,
ing on the length of the HR arms and the fungal species downregulation of the swi6 transcription factor by RNAi
involved (Zhang et al. 2016; Fang et al. 2016; Pohl et al. leads to a decrease in β-glucan and chitin with minor
2016). In this way, a desired mutation can theoretically be growth defects (Lian et al. 2021).
designed and introduced into any gene of interest.
For material use, genetically modified (GM) mush- Mycelium production process and mycelium‑based
room-forming fungi are more acceptable compared to products
edible consumption. In this regard, conventional genetic Classification of mycelium‑based products
modification, as well as genome editing, can be powerful Mushroom mycelium has emerged as a versatile and
tools for molecular breeding. The genes to be disrupted, sustainable material with applications across several
modified or overexpressed should have meaningful traits industries. These applications include mycelium-based
to be molecular breeding targets. Identifying the genes composite, mycelium-based textiles, and many fusion
responsible for cell wall synthesis, growth, and aerial technologies (Fig. 3). Commercialization has been suc-
hypha formation, is indispensable for elucidating cell wall cessful in some product areas (textiles), and many efforts
formation in mushroom-forming fungi and for develop- are underway to commercialize in the areas of high-per-
ing new materials with enhanced properties not found in formance materials (composites and fusion products)
current mycelial materials derived from natural strains. and mass production (leathers and alternative meats).
Innovative applications of mushroom mycelium face sev-
Possible target genes for molecular breeding eral significant challenges that hinder large-scale com-
The cell wall is an ideal target for improving the physical mercialization. A more detailed breakdown by product is
properties of mushroom materials, as it is closely related as follows.
to the stiffness or smoothness of the mycelium mat, as Packaging Mycelium-based composite materials are
discussed in the mushroom composites and packaging being developed as biodegradable alternatives to plas-
section. To dynamically change the cell wall structure, tic and foam packaging. These decompose in as little as
transcriptional regulatory systems of cell wall synthe- 30 days and release nutrients into the soil, making them
sis genes are potential targets. In filamentous fungi, the an eco-friendly option for reducing packaging waste
cell wall integrity (CWI) signaling pathway is known to (Alaneme et al. 2023; Enarevba and Haapala et al. 2023;
be an important transcriptional regulatory system for Abhijith et al. 2018). If the challenges of growth and pro-
cell wall synthesis (Yoshimi et al. 2022). The canonical duction (slow growth rate and scaling limitations) are
CWI pathway is partially conserved in different fungal sufficiently addressed, we expect to see a wider range of
species. It typically comprises cell wall sensors at the cell products coming to the market soon.
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 9 of 21

Mushroom-based materials Substrate (lignocellulosic, etc.)

Mushroom Mushroom byproduct Spent mushroom Agricultural byproduct Forestry byproduct Fibrous materials
Mycelium
(fruiting body) (base, low quality) substrate (SMS) (food waste, etc.) (sawdust, etc.) (cotton, hemp, etc.)

Mycelium-based composite (MBC) Fusion industry (Construction & buildings) Mycelium-based textile (MBT)

Packaging 3D-modeling 3D-printing Inoculum preparation cultivation Substrate sterilization Fiber production Textiling Fashion, leather

Design & Furniture Post-cultivation Mold cultivation Buildings Tiles Insulation panel Mat production Post-processing Wound dressing

Fusion industry (Car seat & leather) Fusion industry (Electronical) Fusion industry (Food & medicine)

Food

Car console Mold cultivation Vinyl cultivation Material production Post-processing Material production Mushroom Mix & Blending

pharmaceutical

Car seat Post-processing Scale-up Bio-circuit board Bio-sensor Wearable device SMS Extract Cosmetics

Fig. 3 Schematic of the production of mycelium-based materials using mushroom mycelium and industrial substrates

Construction and building materials Mycelium com- in plant-based foods since the 1960s. These fungi-based
posites are being researched for use in construction as foods produce 40–52% less pollution compared to live-
insulation panels, building panels, bricks, and sound stock farming (Amara and El-Baky 2023; Souza Filho
panels. Their low density and water resistance properties et al. 2019). Mycelium-based food ingredients must com-
make them promising alternatives in the building indus- pete with traditional food ingredients that have estab-
try (Alaneme et al. 2023; Gou et al. 2021; Joshi et al. 2020; lished supply chains and familiar properties.
Jones et al. 2020). However, the irregular growth patterns Biomedical applications Fungi are being researched for
of mycelium lead to non-uniform thickness and unpre- their potential in synthesizing biomaterials with applica-
dictable properties, making it difficult to achieve stand- tions in medicine and pharmacology (Manan et al. 2021).
ardized products. The lack of a standardized approach to production meth-
Textiles and fashion Mycelium is being used to create ods or material characterization makes it challenging to
leather alternatives for the fashion industry. Mycelium- ensure consistent quality. In addition, the novel nature of
based leather can be dyed and bleached like animal mycelium-based materials may lead to unclear or com-
leather but has a shorter production time of about 5 days plex regulatory pathways for certain applications.
compared to 6–8 weeks for animal leather. It is also bio- Environmental remediation Mycelium is being
degradable and addresses ethical concerns related to explored for its potential in bioremediation and environ-
animal farming (Crawford et al. 2024; Silverman et al. mental cleanup applications (Akpasi et al. 2023). Com-
2020; Bustillos et al. 2020; Cerimi et al. 2019). However, mercialization requires field studies in a wider range of
mycelium-based leathers have relatively low mechani- areas. Additionally, issues related to intellectual property
cal strength, moisture sensitivity and limited thickness, must be addressed. A few companies hold essential pat-
affecting commercial applications such as car seats. ents, restricting knowledge distribution and hindering
Food industry Mycelium is being used to develop meat widespread production.
alternatives and protein sources. Mycoprotein, derived Automotive industry: pressed mycomaterials are being
from the mycelia of Fusarium venenatum, has been used researched for potential use in car parts (Jones et al.
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 10 of 21

2021). Mycelium-based materials have relatively weak techniques depending on the application. Developing
structural properties, limiting their use in load-bearing food, cosmetic, and pharmaceutical ingredients using
applications. Mycelium-based products must compete mycelium requires effectively utilizing edible strains and
with traditional materials including plastics and fiber- spent mushroom substrate (SMS).
reinforced plastics (FRP).
Design and furniture: mycelium is being used to create Culture media and solid substrates
various design objects and furniture pieces, such as lamps Cultivating mushroom mycelium efficiently requires
and chairs (Alemu et al. 2022; Yang et al. 2021). These careful consideration of the culture media and solid
small indoor suites are being commercialized in a variety substrates employed. Mushroom growers have long rec-
of areas, including designer products. One issue is that ognized the importance of optimizing culture condi-
the unique appearance and texture of mycelium materi- tions to enhance mycelial growth and productivity. One
als may not align with traditional consumer preferences. study focused on the entomogenous fungus Ascherso-
Electronics and batteries: the unique properties of liv- nia aleyrodis highlighted the significance of optimizing
ing mycelium networks allow for the propagation of elec- nutritional requirements for both mycelial growth and
trical signals, suggesting potential applications in analog sporulation. (Zhu et al. 2008). When the mycelium of a
electronics and unconventional computing (Mayne et al. mushroom creates a three-dimensional structure and
2023). The integration of mycelium with cellulose-based this structure grows in length, it creates a layer of myce-
materials further enhances the development of biode- lium of a certain thickness, called a mycelium mat. These
gradable batteries, capable of generating power densi- mats can be used as raw materials for mushroom leather
ties suitable for small electronic devices (Danninger et al. and mushroom pulp, and when combined with other
2022). Along with uniform quality control issues, grow- biomass, they form mycelium composites. Culture sub-
ing large volumes of mycelium present logistical difficul- strates are sterilized and cooled in a vinyl bag, inoculated
ties, especially for on-site production. with mushroom hyphae, cultured for a certain period of
These diverse applications showcase the potential of time, and then crushed and put in a container for myce-
mushroom mycelium as a sustainable, biodegradable, and lium mat production and secondary culturing (Gandia
versatile material across multiple industries, offering eco- et al. 2022; Raman et al. 2022). However, this method has
friendly alternatives to traditional materials. Overcoming disadvantages as it is time-consuming due to the need for
these challenges mentioned above will require continued two rounds of culturing and carries a high contamination
research, development of standardized production meth- risk. In addition, establishing optimal culture conditions
ods, and efforts to educate both industry professionals has been attempted using a more effective medium than
and the public about the potential of mycelium-based potato dextrose broth (PDB), which is a medium com-
materials. Collaboration between academia, industry, monly used for culturing mushroom mycelium (Haneef
and regulatory bodies will be crucial in addressing these et al. 2017). This study aimed to find an efficient method
obstacles and unlocking the full potential of mushroom to shorten mycelium mat production time, reduce con-
mycelium in various applications. tamination risk, and improve mycelium mat quality,
Many mycelium-based products (from packaging to thereby increasing productivity and economic value.
electronics) are produced using the solid-state culture Culture media composition plays a crucial role in sup-
process, as shown in Fig. 3. Before the main production porting the growth and metabolism of mushroom myce-
stage, liquid culture and small-scale solid-state culture lium. Techniques such as orthogonal design, response
must be performed to make inoculum and subculture surface design, and uniform design, have demonstrated
materials. To describe the process more comprehensively, effectiveness in optimizing fermentation media, thereby
substrate and culture media must first be dealt with. enhancing fermentation levels efficiency and enzyme
In the case of mycelium-based composite (MBC) and production. This approach can be extended to optimiz-
car console production, 3D modeling is essential to creat- ing culture media for mushroom mycelium cultivation by
ing the formwork for packaging and designing furniture. exploring the optimal concentrations of various nutrients
Some treatments to prevent fire and external contamina- and carbon sources. Recently, a paper was published on
tion are important for construction and building applica- the importance of disaccharides as a carbon source in
tions. Mycelium-based textiles (MBT) and leather require mushroom mycelium cultures (Nussbaum et al. 2023).
the application of chemical processes such as tanning, Malt concentration in solid substrate influences mycelial
dyeing, and surface coating used in conventional leather growth and network connectivity in Ganoderma species,
processes. For electronic applications, this is comple- impacting radial growth kinetics, mycelium density, and
mented by cultivation techniques to fine-tune the shape mechanical characteristics like Young’s modulus. In addi-
and density of the hyphae, as well as post-processing tion, the effect of agar, another carbohydrate component,
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 11 of 21

is also being studied. In the past, agar was not consid- available from some companies, they are not compara-
ered an energy or carbon source, but it has recently ble to traditional leather in terms of texture or durabil-
been recognized as an important factor in the growth of ity. Furthermore, there are still too many affordability
hyphae and mycelium. When the agar concentration was hurdles to overcome. As a reflection of this, some mush-
increased from 1.5% to 6.0% agar, the mycelium density room leather companies in the US have ceased produc-
and hyphal width increased by 32.3% and 63.6%, respec- tion (Chan and Webb 2023). Studies are actively being
tively. The implications of these findings will allow for conducted to meet the industrial standard of leather and
the advancement and tuning of fungi-based materials, pulp through physical and chemical treatment processes
particularly for the application of sustainable textiles and (Appels et al. 2020; Raman et al. 2022).
fine particulate filters (Hotz et al. 2023). In the future, the Mushroom pulp, often referred to as mycelium pulp,
effects of different carbohydrate components, including is an innovative bio-based material derived from the
maltose and agar, should be studied more closely. Table 1 mycelial structure of mushrooms. The production of
summarizes media for strains stock, liquid media for pre- mycelium pulp, achieved through innovative extrac-
pare mycelium inoculum and solid media for mycelium tion methods from fruiting bodies, retains the structural
production. integrity of the mycelium, resulting in a versatile material
Solid substrates also present an important factor in with excellent deformability suitable for diverse applica-
the cultivation of mushroom mycelium. Using agricul- tions, including packaging materials, textiles, and even
tural waste or byproducts as solid substrates offers a building materials (Nakauchi et al. 2023). Additionally,
cost-effective and sustainable solution, while potentially the cultivation of mycelium on paste substrates has been
influencing the growth characteristics and secondary explored to improve nutrient availability and scalabil-
metabolite production of the mycelium (Ángeles-Argáiz ity, further supporting the feasibility of mycelium-based
et al. 2020). Most agricultural residues rich in lignocellu- products (Crawford et al. 2024). However, challenges
lose, such as wheat straw, rice straw, corn cobs, and sugar remain in replicating the full properties of leather and
cane, serve as ideal substrates for fungal growth due to optimizing processing techniques, indicating a need for
cellulose, hemicellulose, and lignin composition. These continued research in this field (Kniep et al. 2024; Raman
materials typically contain 30–50% cellulose, 20–30% et al. 2022). Mycelium offers a sustainable alternative to
hemicellulose, and 5 to 30% lignin. Predictions suggest conventional materials, aligning with eco-friendly design
that global waste production could reach 2.59 billion tons principles.
by 2030 (Awogbemi and Von Kallon 2022). Therefore,
the efficient utilization of various cellulosic by-products Mushroom composites and packaging (plastic alternatives)
generated globally as solid media for raw materials is Among the materials, the most widely used material is
expected to significantly contribute to carbon reduction plastic. There are many types of plastic and it is the most
and the economic production of mushroom mycelium. commonly encountered material in our daily life. How-
Representative substrates for mycelium composite pro- ever, since it is a petroleum-based material, even if the
duction reported so far are shown in Table 1. oil depletion problem is solved, the environmental prob-
The optimization of culture media and solid substrates lem remains a big issue. Mushroom mycelium presents
is crucial for enhancing the efficiency and productivity a promising alternative to traditional plastic packaging,
of mushroom mycelium cultivation. By drawing upon leveraging its biodegradable and sustainable properties.
insights from related fields, such as mushroom research Mycelium, derived from agricultural waste, forms robust,
and fermentation medium optimization, researchers and self-assembling materials that can replace polystyrene in
growers can develop innovative strategies to unlock the packaging applications due to its environmental friendli-
full potential of mushroom mycelium production. ness and ability to degrade naturally (Patel and Sharma
2023). Research highlights the versatility of mycelium
Mushroom leather and pulp composites, which exhibit high stability, flexibility, and
Mushroom leather and pulp production using myce- mechanical strength, making them suitable for various
lium presents a promising avenue for sustainable mate- uses, including insulation and packaging (Verma et al.
rial development. Mycelium-based leather, derived from 2023; Zhang et al. 2023). Innovative approaches, such
various fungi, has shown mechanical properties com- as glycerol treatment, have enhanced the toughness and
parable to traditional leather, particularly when rein- hydrophilicity of mycelium films, further improving their
forced with substrates like needle felts or woven fabrics, applicability in packaging (Amobonye et al. 2023). Addi-
enhancing tensile strength beyond that of synthetic tionally, studies on mycelium composites derived from
alternatives (Kniep et al. 2024; Raman et al. 2022). While hemp shives indicate complete biodegradability within
various mushroom leathers have been researched and are 12 weeks, although challenges like high water absorption
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 12 of 21

Table 1 Summary of culture medium and solid substrate for the production of mushroom mycelium
Strain Medium for strain Liquid medium to Solid medium for Substrate for References
stock prepare mycelium mycelium mat mycelium composite
inoculum production

Abortiporus biennis Malt extract glucose Low nutrient media – Wheat bran, Wheat Balaes et al. (2023)
agar straw, Coconut husk,
Broadleaves sawdust
Ceriporia lacerata Potato dextrose agar Potato dextrose – Soybean straw Shao et al. (2016)
medium
Fomitopsis pinicola Malt extract peptone 1% Sugar, 1% Malt, 0.2% Woven fabrics, Woven Woven fabrics, Woven Kniep et al. (2024)
agar Yeast felts felts
Fomitella fraxinea Potato dextrose agar Yeast malt peptone Sawdust substrate – Raman et al. (2022)
medium
Potato dextrose agar Potato dextrose – Sawdust substrate Bae et al. (2021)
medium
Fomes fomentarius Malt extract agar Malt extract medium – Hemp shives Pohl et al. (2022)
Malt extract glucose Low nutrient media – Wheat bran, Wheat Balaeș et al. (2023)
agar straw, Coconut husk,
Broadleaves sawdust,
Ganoderma lucidum – – – Bamboo fiber substrate Soh et al. (2020)
Potato dextrose agar Potato dextrose Microcrystalline cel‑ – Haneef et al. (2017)
medium lulose
– – – Red oak sawdust, Cot‑ Lingam et al. (2023)
ton carpel
– – – Beech sawdust, cotton Vasatko et al. (2022)
fiber, soy silk fiber,
wheat bran
Grain mixture Grain mixture – P. eryngii Spent Schritt et al. (2021)
mushroom substrate
with wheat bran, wheat
meal, oil seed
Grain mixture Hemp hurds + maple – Hemp fibers, Hemp Özdemir et al. (2022)
veneers hurds
Pine wood sawdust
and shavings
Ga. resinaceum – – – Hemp shives & Soybean Adamatzky and Gandia
hulls (2022)
Dark Malt extract agar Dark Malt extract – Wheat straw Xing et al. (2018)
medium
Letinus sajor-caju Wood Potato dextrose – Corn husk, Wood saw‑ Teeraphantuvat et al.
medium Sorghum grain dust, 5% Rice bran, 1% (2024)
Calcium carbonate, 2%
Calcium sulfate, 0.2%
Sodium sulfate
Trametes multicolor – – – Rapeseed straw, Beech Appels et al. (2019)
sawdust
Tr. versicolor Potato dextrose agar Potato dextrose – Sawdust substrate Bae et al. (2021)
medium
Grain mixture Grain mixture Bacterial cellu‑ – Elsacker et al. (2021)
lose + hemp fibres
Malt extract agar Malt extract agar + ster‑ – Betula alleghaniensis Sun et al. (2022a, b)
ile corn steep liquor Britt. wood veneers
Malt extract glucose Low nutrient media – Wheat bran, Wheat Balaeș et al. (2023)
agar straw, Coconut husk,
Broadleaves sawdust
Tr. orientalis Potato dextrose agar Yeast malt medium Sawdust substrate – Jeong et al. (2023)
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 13 of 21

Table 1 (continued)
Strain Medium for strain Liquid medium to Solid medium for Substrate for References
stock prepare mycelium mycelium mat mycelium composite
inoculum production

Pleurotus ostreatus – – – Cotton Appels et al. (2019)

Potato dextrose Potato dextrose broth Microcrystalline cel‑ – Haneef et al (2017)
medium lulose
Potato dextrose agar Potato dextrose – Sugar Cane, Sawdust,
broth + 1% cellulose wheat bran,
– – – Straw, Sawdust, Lingam et al. (2023)
Bagasse + saw‑
dust + wheat bran
– Glucose agar – Rubber wood saw‑ Shakir et al. (2020)
dust + corn grain
– – Hemp bedding sub‑ – Adamatzky and Gandia
strate (2021)
Pl. sanguineus Potato dextrose agar Potato dextrose – Coconut pow‑ Gou et al. (2021)
medium der + wheat bran
Pl. eryngii – – – Sawdust substrate Silverman et al. (2020)
Malt extract peptone 1% Sugar, 1% Malt, 0.2% Woven fabrics Woven Woven fabrics Woven Kniep et al. (2024)
agar Yeast felts felts

and mold resistance remain (Loris et al. 2022). While fatty acid profiles when cultivated on food industry
mycelium-based materials show great potential, ongo- byproducts like tempeh residual water (Wikandari et al.
ing research is necessary to optimize their properties for 2023). Additionally, various fungal species, such as
broader commercial use. To meet the industrial stand- Fusarium venenatum and Le. edodes, have been identi-
ards of plastic products including packaging, the devel- fied as suitable candidates for meat replacement due to
opment of lightweight materials with high stiffness is their nutritional profiles and rapid growth rates (Kok Kee
essential. In addition, research on optimizing production et al. 2022). Moreover, edible mycelium can function as a
systems for commercializing mycelium-based materials support structure in cultivated meat production, enhanc-
is ongoing. Despite this high possibility, mass production ing cell proliferation and differentiation, which is crucial
using mushroom mycelium has not yet been reported in for developing meat-like textures (Ogawa et al. 2024).
the paper but strategies for growing large-scale mycelium Despite challenges in mycelium productivity and sensory
structures were suggested (Dessi-Olive 2022). Several acceptance, advancements in culture medium optimiza-
companies continue to promote the mass production of tion show potential for improving these aspects (Majum-
mushroom composites (Bearne 2023). der et al. 2024). Overall, leveraging mushroom mycelium
could significantly contribute to sustainable alternative
Alternative (artificial) meat protein sources (Holt et al. 2024).
The representative of alternative livestock products is
artificial meat. Artificial meat can be divided into two Construction materials
main methods: one involves mimicking the taste and Mycelium composites are emerging as a promising sus-
texture of meat by processing plant materials, while the tainable alternative in the construction industry due to
other involves culturing animal cells in specific areas their eco-friendly properties and potential for innova-
and conditions. These artificial meats have some advan- tion. These materials are derived from the vegetative part
tages over conventional meat in terms of safety and ani- of fungi, known as mycelium, and can be combined with
mal cruelty issues. Alternative meat production using lignocellulosic materials to form mycelium-based com-
mushroom mycelium presents a promising avenue for posites (MBCs). These composites are characterized by
addressing global protein demands while mitigating envi- their low density, elasticity, and cost-effectiveness, mak-
ronmental impacts. Research indicates that filamentous ing them suitable for various applications in construction
fungi, particularly mycoprotein derived from mycelium, and other industries (Madusanka et al. 2024). Mycelium
can serve as nutritious meat substitutes, with studies composites offer several advantages over traditional
showing high protein content (19.4% w/w) and beneficial materials. They are biodegradable, contribute to zero
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 14 of 21

waste during production, and can be tailored through mapping of electrical signals due to the substrate’s non-
genetic and biochemical modifications to enhance their linear electrical characteristics, which allows it to func-
properties. The versatility of MBCs allows them to be tion as a living electronic component (Roberts and
used in material-driven design (MDD) approaches, Adamatzky 2022). Mycelia, the branching, web-like root
which has the potential to foster innovative applica- structure of fungi, can act as conductors and electronic
tions in construction and other industries (Madusanka components (Adamatzky and Gandia 2021). This capa-
et al. 2024). Despite their potential, mycelium compos- bility is being investigated by researchers, such as those
ites face several challenges. The production process can at the Unconventional Computing Laboratory, who are
have a significant environmental impact, particularly in developing fungal computing (so called mushroom com-
regions where fossil fuels are the primary energy source. puters) and electronics by leveraging the action poten-
For example, in South Africa, the high electricity con- tial-like spikes produced by fungi, similar to neuronal
sumption required for production can make mycelium spikes in the brain (Hu 2023). These spikes can be used
composites less sustainable than conventional materials to represent binary data (zeroes and ones) and to imple-
like concrete. Additionally, the shorter lifespan of these ment basic logical and electronic circuits (Adamatzky
composites necessitates frequent replacements, further and Gandia 2021). Fungal mycelium has been shown to
increasing their environmental footprint (Akromah et al. increase conductivity and communication speed when
2024). stimulated at two separate points, which allows for the
Ongoing research is focused on overcoming these establishment of memory, akin to how brain cells form
challenges by improving the biological characteristics of habits. Different geometries of mycelium can compute
mycelium through bio-fabrication procedures and opti- various logical functions, which can be mapped based on
mizing the production process (Alaneme et al. 2023). its electrical responses (Phillips et al. 2024). Nowadays,
Researchers are also conducting life cycle assessments taking advantage of fungal mycelia’s natural light sensitiv-
to understand the environmental impacts better and to ity, Mishra et al. (2024) developed an electrical interface
develop strategies for reducing them, such as using alter- to both house the mycelia and measure their electrophys-
native energy sources (Volk et al. 2024; Alaux et al. 2024; iological action potentials. The following are potential
Enarevba and Haapala et al. 2023; Williams et al. 2022). applications:
Collaborative efforts in material science and fungal bio- Environmental sensors Fungal networks can be used
technology are encouraged to address these issues and as large-scale environmental sensors. They can monitor
unlock the full potential of mycelium composites for sus- data flows and provide insights into ecosystem changes
tainable construction. While mycelium composites hold by interpreting the electrical signals used by fungi to pro-
great promise for sustainable construction, their envi- cess information (Dehshibi et al. 2021).
ronmental impact and production challenges must be Biodegradable electronics: fungal machines offer the
carefully managed. Continued research and innovation potential for creating self-assembling, self-repairing, and
are essential to enhance their viability as a green building biodegradable electronic devices. These could be used
material. in situations where traditional electronics are not feasible
or environmentally friendly (Danninger et al. 2022).
Electronics Wearable technology researchers are exploring the use
The use of mushroom mycelium in biocomputing and of mycelium networks in wearable technology. Fungal
biosensing is an emerging field that explores the poten- skins have shown potential as touch sensors in robot-
tial of exploring fungi as computing devices and sen- ics and can sense light, making them suitable for various
sors (Fukasawa et al. 2024; Phillips et al. 2024; Mayne wearable applications (Adamatzky et al. 2021).
et al. 2023; Danninger et al. 2022). Filamentous fungi, Structural and building materials fungal mycelium can
such as those found in mycelial networks, have long create structural substrates for buildings that self-grow,
been recognized for their ecological and biotechno- build, and repair themselves. These structures could
logical importance, yet synthetic biologists have his- include embedded sensorial elements for environmental
torically overlooked them for electronic application. The monitoring (McGaw et al. 2022).
increased complexity of their genomes and the lack of Optimization and artificial intelligence: mushroom
molecular tools have slowed the pace of innovation in computers can solve complex problems in fields like
filamentous fungi engineering. However, recent advance- artificial intelligence and optimization by leveraging the
ments in synthetic biology have unlocked the potential adaptability and complexity of living fungal networks
of these remarkable organisms, paving the way for their (Wainaina and Taherzadeh 2023).
integration into the world of biocomputing and biosens- These applications highlight the unique capabilities of
ing (Jo et al. 2023). The mycelium is capable of nontrivial fungal computing, particularly in areas where traditional
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 15 of 21

computing technologies may face limitations. However, compound is primarily used as the positive electrode
it is important to note that fungal computing is still in material. Mushroom mycelium has been explored as a
the research phase, and further development is needed sustainable and flexible material for the fabrication of
to realize its potential in everyday devices. In addition, microsupercapacitors, which can serve as an alternative
applications in this field highlight further beneficial to traditional lithium-ion batteries (Coelho et al. 2023).
mycelial properties, differing those of the aforemen- These mushroom-based microsupercapacitors exhibit
tioned mycelium materials. desirable characteristics such as high-power densities,
long life cycles, and fast charge–discharge rates, making
Capacitors and batteries them a promising option for various portable and wear-
Capacitors and batteries are essential components in able technologies. Mushroom mycelium contributes to
modern electronic devices, serving different yet comple- sustainable battery production through its application
mentary functions. Capacitors are essential components as a biodegradable substrate and as a source of bioactive
in electronic circuits, playing a crucial role in energy materials. Research demonstrates that fungal mycelium
storage, filtering, and signal coupling. In recent years, the skins can be processed into flexible electronic devices,
development of electrochemical capacitors, also known exhibiting high conductivity and durability, essential
as supercapacitors, has garnered significant attention due for battery applications (Danninger et al. 2022). These
to their superior power capabilities and exceptional life mycelium-based batteries have approximately 3.8 mAh
cycle (Zhao and Burke 2021). One approach to enhanc- ­cm−2 capacities, effectively powering small electronic
ing supercapacitor performance is exploring new mate- devices like sensors (Danninger et al. 2022). Additionally,
rials, such as fungal-derived compounds (Szacilowski mycelium residues from fermentation processes, such as
et al. 2023). Recent trends in fungal capacitor research those from Aspergillus niger, can be converted into bio-
highlight the innovative use of mycelium and fungal- char, which serves as an anode material for lithium-ion
derived materials in energy storage applications. Studies batteries. This approach utilizes waste and enhances bat-
indicate that mycelium, particularly from species like Pl. tery performance due to the nitrogen and silicon content
ostreatus, exhibits promising capacitive properties, with in the mycelium (Gu et al. 2023). Mushroom-derived
capacitance values in the hundreds of picofarads and a biomasses and mycelia are being extensively studied
voltage-dependent pseudocapacitance reaching hundreds as anode materials for batteries (Li et al. 2024; Xu et al.
of microfarads. This suggests potential for mycelium 2023; Han et al. 2020; Tang et al. 2016; Campbell et al.
as an organic alternative in capacitor technology, espe- 2015). Research shows that the anode material of mush-
cially in dense arrays for charge storage (Szacilowski et al. room-based components can omit the activation pro-
2023). Additionally, research on biomass-derived fungal cess, and is environmentally friendly as the use of various
carbons, such as those from Ag. bisporus and Pl. eryngii, anode activation and coating reagents. These findings
revealed their high surface areas and specific capacitance, collectively highlight mycelium’s potential in advancing
making them suitable for supercapacitor applications. sustainable battery production while addressing environ-
These materials can be tuned for optimal performance mental concerns associated with electronic waste.
through variations in growth conditions and species
selection, achieving specific capacitance values exceeding Mycelium‑production bioprocess
those of traditional carbon materials (Jones et al. 2024). In the last decade, great progress has been made in
Furthermore, activated carbon derived from black Asper- mushroom mycelium research, from basic research to
gillus demonstrates exceptional gravimetric capacitance various applied materials. However, for mushroom myce-
and cycling stability, indicating a sustainable pathway lium research to make significant industrial progress,
for high-performance supercapacitors (Yang et al. 2021). large-scale production and corresponding bioprocessing
Overall, the integration of fungal materials into capacitor systems are urgently needed. In particular, we are con-
technology represents a significant advancement in sus- vinced that the large-scale production and utilization of
tainable energy storage solutions. mushroom mycelium will increase dramatically if even a
The relationship between these two energy storage partial continuous process is possible, as the current pro-
devices has become increasingly important, especially cess is still batch-based.
with the rise of emerging technologies such as lithium- Mushroom cultivation is a unique microbial process
ion batteries and their potential applications. (Coelho that can effectively bioconvert a wide range of plant
et al. 2023; Docimo et al. 2014; Porcarelli et al. 2012). polymers, including lignin, cellulose, and hemicellu-
Lithium-ion batteries are largely composed of a posi- lose (Lakhanpal 2002). This process produces edible and
tive electrode, a negative electrode, a separator, and an nutritious mushrooms and provides an efficient and eco-
electrolyte. A lithium-transition metal oxide-based nomically viable method for valorizing lignocellulosic
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 16 of 21

waste materials. (Kumar et al. 2018). While there are regulates the thickness and growth direction of the myce-
some similarities to conventional mushroom cultivation, lium, is a challenge that needs to be addressed. In addi-
the cultivation of mushroom mycelium in this article is tion, as the growth direction, shape, and thickness of the
largely different. Mycelium cultivation involves growing mycelium are varied, the mycelium content may differ
only mycelium quickly and in large quantities while main- depending on the sample collection location. Therefore,
taining conditions that prevent the production of fruiting it is unlikely that the same amount of mycelium will be
bodies. The waste substrate can be converted into bio- contained in each individual product, regardless of the
energy, biofertilizer, and biocompost, thereby reducing type of composite material manufactured. This makes
the environmental impact of these waste streams. (Wan commercializing difficult as it is challenging to obtain the
Mahari et al. 2020) Furthermore, cultivation is labor- same quality amongst all products. To overcome these
intensive and can provide employment opportunities, difficulties, the role of biochemical engineering research-
particularly in rural areas. (Kumar et al. 2018). ers will be crucial in the future. First, it is necessary to
Mushroom mycelium grows radially, and becomes secure culture technology that can produce mycelium
entangled to form a biofilm (mycelium mat). In addition, in a short period, develop a device that ensures uniform
the mycelium exchanges oxygen and carbon dioxide nec- material quality by controlling mycelium growth, and
essary for proliferation to form aerial hyphae. The struc- improve physical and biochemical properties through
ture in which the thickness and density are increased mixing with various materials. Additionally, cutting tech-
through the growth of biofilms and aerial mycelium niques suited to the user are required. If these technical
is called a mushroom mycelium mat. To produce a difficulties are overcome, the transition from existing
mushroom mycelium mat of high quality, optimiza- plastic-based composite materials to fungal mycelium-
tion of the production process, including raw materials, based composite materials is expected to be successfully
unit operations, culture bioreactors, process controls, achieved.
and economic analysis, is essential. The final mush-
room mycelium products generally have lower physical Conclusion
strength compared to industrial counterparts. Therefore, Most of the materials derived from mushroom mycelium
to meet the industrial standards of each product various discussed in this review are replacements for existing
methods such as plasticization, cross-linking, pressing, products, meaning they must meet various performance
and coating processes are employed (Chan et al. 2021; standards in each material field. Therefore, new stand-
Manan et al. 2021; Yang et al. 2021; Raman et al. 2022; ards and legislation are essential for commercializing a
Elsacker et al. 2023; Teeraphantuvat et al. 2024). broader range of mushroom mycelium products, and
Both batch and continuous processes can be explored organic integration with existing production processes is
for large-scale bioprocessing of mushroom mycelium necessary for developing new industrial products.
production. In a batch process, the growth substrate is Although mushroom mycelium products have a low
inoculated with mushroom mycelium and incubated carbon footprint, the energy requirements for their pro-
under controlled conditions until the desired yield is duction must be addressed. They also require significant
achieved. In contrast, a continuous process involves the investments in research, development, and production
continuous feeding of growth substrate and harvesting facilities.
the produced mycelium, allowing for a more efficient and Finally, we need to solve the challenges posed by fickle
scalable production system. For large-scale production consumers and greenwashing in many parts of the indus-
of mycelium, culture substrates are sterilized and cooled try to ensure that truly sustainable products can remain
in a vinyl bag, inoculated with mushroom hyphae, cul- competitive. Academic research is essential to close the
tured for a certain period, and then crushed and placed gap between research and industrialization, and we sug-
in a container for mat production and secondary cultur- gest that some companies make their production knowl-
ing (Gandia et al. 2022; Raman et al. 2022). In the future, edge publicity available.
automation facilities will be adopted to develop an effi-
Abbreviations
cient production process that shortens the time required POD Peroxidase
for high-quality mushroom mycelium mat, reduces the GH Glucoside hydrolase
possibility of contamination, and improves the quality to SEM Scanning electron microscope
EPS Extracellular polymeric matrix
increase productivity and cost-efficiency. CHS Chitin synthase
There is a possibility that mycelium-based materials GMO Genetically modified organism
will fail to be commercialized. The most challenging part UV Ultraviolet
RAPD Random amplification of polymorphic DNA
in the commercialization of mycelium-based materials is ATMT Agrobacterium tumefaciens-mediated transformation
product homogeneity. The issue of process control, which NHEJ Non-homologous end joining
Shin et al. Bioresources and Bioprocessing (2025) 12:3 Page 17 of 21

DSB DNA double-strand break Adamatzky A, Nikolaidou A, Gandia A, Chiolerio A, Dehshib MM (2021) Reac‑
gRNA Guide RNA tive fungal wearable. Biosystems 199:104304. https://​doi.​org/​10.​1016/j.​
HR arm Homologous arm sequences biosy​stems.​2020.​104304
HDR Homology-direct repair Aiduang W, Jatuwong K, Luangharn T, Jinanukul P, Thamjaree W, Teeraphantu‑
GM Genetically modified vat T, Waroonkun T, Lumyong S (2024) A review delving into the factors
CWI Cell wall integrity influencing mycelium-based green composites (MBCs) production and
GEF Guanine nucleotide exchange factors their properties for long-term sustainability targets. Biomimetics 9:337.
MPK Mitogen-activated protein kinase https://​doi.​org/​10.​3390/​biomi​metic​s9060​337
PKC Protein kinase C Akpasi SO, Anekwe IMS, Tetteh EK, Amune UO, Shoyiga HO, Mahlangu TP,
MBC Mycelium-based composite Kiambi SL (2023) Mycoremediation as a potentially promising technol‑
MBT Mycelium-based textile ogy: current status and prospects—a review. Appl Sci 13:4978. https://​
SMS Spent mushroom substrate doi.​org/​10.​3390/​app13​084978
PDB Potato dextrose broth Akromah S, Chandarana N, Rowlandson JL, Eichhorn SJ (2024) Potential envi‑
MDD Material-driven design ronmental impact of mycelium composites on African communities.
Sci Rep 14:11867. https://​doi.​org/​10.​1038/​s41598-​024-​62561-7
Acknowledgements Alaneme KK, Anaele JU, Oke TM, Kareem SA, Adediran M, Ajibuwa OA,
The authors would like to acknowledge the financial support provided by the Anabaranze YO (2023) Mycelium based composites: a review of their
Rural Development Administration of Korea (RDA). bio-fabrication procedures, material properties and potential for green
building and construction applications. Alex Eng J 83:234–250. https://​
Author contributions doi.​org/​10.​1016/j.​aej.​2023.​10.​012
All authors performed the literature survey and contributed to the writing and Alaux N, Vašatko H, Maierhofer D, Saade MRM, Stavric M, Passer A (2024)
final editing of the paper. Hyun-Jae Shin and Yoichi Honda designed the paper Environmental potential of fungal insulation: a prospective life cycle
and supervised the writing. assessment of mycelium-based composites. Int J Life Cycle Assess
29:255–272. https://​doi.​org/​10.​1007/​s11367-​023-​02243-0
Funding Alemu D, Tafesse M, Mondal AK (2022) Mycelium-based composite: the future
This work was supported under the framework of international coopera‑ sustainable biomaterial. Int J Biomater 12:8401528. https://​doi.​org/​10.​
tion program managed by the National Research Foundation of Korea 1155/​2022/​84015​28
(2023K2A9A2A08000176, FY2023 to HS). Bilateral Program managed by the Amara AA, El-Baky NA (2023) Fungi as a source of edible proteins and animal
Japan Society for the Promotion of Science [JPJSBP 120238807 to M. K.] and feed. J Fungi 9:73. https://​doi.​org/​10.​3390/​jof90​10073
the Institute for Fermentation, Osaka, Japan [K-2019–002 to M K]. Amobonye A, Lalung J, Awasthi MK, Pillai S (2023) Fungal mycelium as leather
alternative: a sustainable biogenic material for the fashion industry.
Availability of data and materials Sustain Mater Technol 38:e00724. https://​doi.​org/​10.​1016/j.​susmat.​
Data will be made available upon request. 2023.​e00724
Ángeles-Argáiz R, Carmona-Reyes IA, Quintero-Corrales C, Medina-Macías
FJ, Blancas-Cabrera A, Valdez-Cruz NA, Ulloa M, Trujillo-Roldán MA,
Declarations Garibay-Orijel R (2020) From field sampling to pneumatic bioreac‑
tor mycelia production of the ectomycorrhizal mushroom Laccaria
Ethics approval and consent to participate trichodermophora. Fungal Biol 124:205–218. https://​doi.​org/​10.​1016/j.​
Not applicable. funbio.​2020.​02.​003
Antinori ME, Ceseracciu L, Mancini G, Heredia-Guerrero JA, Athanassiou A
Consent for publication (2020) Fine-tuning of physicochemical properties and growth dynam‑
All authors read and approved the manuscript for publication. ics of mycelium-based materials. ACS Appl Bio Mater 3:1044–1051.
https://​doi.​org/​10.​1021/​acsabm.​9b010​31
Competing interests Appels FVW, Camere S, Montalti M, Karana E, Jansen KMB, Dijksterhuis
The authors declare no conflict of interest. J, Krijgsheld P, Wosten HAB (2019) Fabrication factors influencing
mechanical, moisture- and water-related properties of mycelium-based
Author details composites. Mater des 161:64–71. https://​doi.​org/​10.​1016/j.​matdes.​
1
Department of Biochemical Engineering, Chosun University, Gwangju, 2018.​11.​027
Republic of Korea. 2 Department of Bio and Medical Big Data (BK4 Program) Appels FVW, van den Brandhof JG, Dijksterhuis J, de Kort GW, Wosten HAB
and Research Institute of Life Sciences, Gyeongsang National University, (2020) Fungal mycelium classified in different material families based
Jinju, Republic of Korea. 3 Laboratory of Environmental Interface Technology on glycerol treatment. Commun Biol 3:334. https://​doi.​org/​10.​1038/​
of Filamentous Fungi, Graduate School of Agriculture, Kyoto University, Kyoto, s42003-​020-​1064-4
Japan. 4 Laboratory of Forest Biochemistry, Graduate School of Agriculture, Arantes V, Goodell B (2014) Current understanding of brown-rot fungal
Kyoto University, Kyoto, Japan. biodegradation mechanisms a review Deterioration and protection
of sustainable biomaterials. In: Schultz P (ed) ACS Symposium Series.
Received: 28 August 2024 Accepted: 23 December 2024 American Chemical Society, Washington
Ariyani NR, Wijayanti SP, Putra NG, Kusumawati MB, Setiawan AAR, Isharyadi
F, Widyastuti N, Djarot IN, Handayani T (2024) Mycelium-based leather
potential for sustainability: a review. AIP Conf Proc 3001:030012. https://​
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